畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (3): 380-384.doi: 10.11843/j.issn.0366-6964.2014.03.006

• 遗传繁育 • 上一篇    下一篇

LIAS基因微环表达载体的构建

高会贞,王晓婷,郭豫杰,韩立强,鲁维飞,杨国宇*   

  1. (河南农业大学 农业部动物生化与营养重点开放实验室,郑州 450002)
  • 收稿日期:2013-07-03 出版日期:2014-03-23 发布日期:2014-03-23
  • 通讯作者: 杨国宇,教授,博士,主要从事动物生物化学研究,E-mail: haubiochem @163.com
  • 作者简介:高会贞(1987-),女,河南新乡人,硕士生,主要从事动物生物化学研究,E-mail:gaohuizhen0716@163.com
  • 基金资助:

    农业部“引进国际先进农业科学技术”(948)重点项目(2011-G35)

Construction of Minicircle Vector Carrying Porcine LIAS Gene

GAO Hui-zhen,WANG Xiao-ting,GUO Yu-jie,HAN Li-qiang,LU Wei-fei,YANG Guo-yu*   

  1. (Key Laboratory of Animal Biochemisty and Nutrition,Ministry of Agriculture,Henan Agricultural University,Zhengzhou 450002, China)
  • Received:2013-07-03 Online:2014-03-23 Published:2014-03-23

摘要:

为了构建猪LIAS基因长期高效的表达载体,本研究通过RT-PCR方法从猪肌肉组织中克隆了LIAS基因片段,并将该基因片段与微环表达载体minicircle进行连接,构建成微环载体minicircle-LIAS,然后转染HeLa细胞,检测其在HeLa细胞中绿色荧光蛋白的持续表达时间。结果表明,本研究克隆了猪LIAS基因,该基因长度为1 119 bp,并将该基因成功构建到了微环表达载体minicircle上;转染HeLa细胞对绿色荧光蛋白持续表达时间的观察显示,构建的微环表达载体minicircle-LIAS中绿色荧光蛋白的持续表达时间明显高于常用的pEGFP-N1质粒,为进一步阐明LIAS合成的生化过程及功能调控提供了新思路和试验材料。

Abstract:

In order to construct a long-term and high-level transgene expression recombinant vector,LIAS gene was cloned from the muscle of porcine by RT-PCR and inserted into minicircle plasmid.The minicircle-LIAS vector was transfected into HeLa cells to investigate the duration of GFP expression.The results showed that the LIAS gene which was 1 119 bp length was completely cloned and inserted into minicircle plasmid.The vector was transfected into HeLa cells and the duration of GFP expression of recombinant minicircle-LIAS vector was obviously higher than that of pEGFP-N1 plasmid.This result provides a new thought and experimental material for the process of LIAS biological synthesis and function regulation.

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